Ateneo - UPO

The course will provide the basics of the instrumental analytical techniques most used in the clinical laboratory (liquid chromatography, gas chromatography, electrophoretic techniques, elementary analysis, etc.), together with the most used pre-treatment and sample analysis methods in the clinical laboratory. The performance indexes of an analytical method (LOD, LOQ, recovery, dynamic and linear range, robustness, measurement uncertainty, accuracy, repeatability, etc.) will also be addressed. Finally, the principles and applications of chemometric methods of identification of biomarkers will be presented.

Material from the teacher

The course aims to provide students with knowledge of the most widespread analytical instrumentation (liquid chromatography, gas chromatography, electrophoretic techniques, elementary analysis) and on parameters to evaluate the performance of an analytical method. Knowledge is also provided about the sample pretreatment methods for the clinical laboratory and the fundamentals of the analytical methods used for the determination of blood and urinary parameters.
Finally, knowledge of multivariate analytical methods for the identification of pathology biomarkers will be provided.
The student will acquire critical sense skills, refining the ability to choose the best experimental strategy to solve a given analytical problem.
Communication skills will also be stimulated through an appropriate vocabulary.

None

Lectures, classroom discussions with students

Learning will be evaluated through classroom discussions with the students.

Oral exam consisting of three open questions: one on the theoretical and practical aspects of the analytical techniques seen in class; one on the determination of analytical parameters in biological samples; one on biomarker identification methods.

Part I - Instrumental analysis techniques.
Introduction to spectroscopic techniques, electromagnetic radiation, absorption, emission, fluorescence, phosphorescence.
Molecular UV-visible absorption: principle, instrumentation, applications.
Elementary techniques: introduction, flame and furnace atomic absorption, instrumentation, interferences, correction methods, applications; inductive coupling plasma, instrumentation, applications.
Chromatographic techniques: introduction, plate and velocity theory, liquid chromatography (mechanisms, instrumentation, applications), gas chromatography (mechanisms, instrumentation, applications).
Electrophoretic techniques: capillary electrophoresis, electrophoretic and non-electrophoretic immunoprecipitation techniques, gel techniques, applications.
Performance indexes of an analytical method (LOD, LOQ, recovery, dynamic and linear range, robustness, measurement uncertainty, accuracy, repeatability, etc.).
Part II - Analysis of biological samples.
Sample collection and storage methods. Analytical determinations in the chemical-clinical laboratory: electrolyte balance, trace elements, calcium, magnesium and major cations, lipids and lipoproteins, protein metabolism, bilirubin and molecules related to jaundice, carbohydrate metabolism, balance of bone remodeling, enzymes, circulating markers, hormones, urine tests.
Part III - Biomarkers.
Introduction to multivariate methods. Monovariate methods: T-student test. Multivariate methods: correlation, data pre-treatment, analysis of the main components, classification methods, the problem of identifying biomarkers, confusion matrix, sensitivity, specificity, precision, accuracy, non-error-rate%, ROC curve.

Knowledge and understanding
- knowledge of the theoretical and theoretical / practical bases of the most modern analysis techniques (chromatography, elementary methods, UV-visible spectroscopy, electrophoretic techniques, mass spectrometry)
- knowledge of pre-treatment methods and analysis of biological samples
- knowledge of the principles of multivariate analysis and of the main methods of pattern recognition and classification

Ability to apply knowledge and understanding
- know how to compare different analytical techniques
- know how to choose the best method for the determination of analytical parameters in biological samples
- to be able to comment on a ROC curve and compare screening methods and confirmatory methods

Communication skills
- acquire and know how to use an appropriate vocabulary in relation to the topics addressed in the course

Autonomy of judgment
- ability to choose between different analytical techniques for the same purpose